西北地区530例临床患者CYP2C19遗传多态性分析

目的采用DNA微阵列芯片法对西北地区人群等位基因CYP2C19*2和*3进行基因多态性检测,以了解本地区人群CYP2C19基因型。方法在完成PCR扩增获得目标片段后,运用DNA微阵列芯片法,通过具有位点特异性的寡核苷酸探针与目标序列的杂交及显色,判读基因多态性类型。结果在530份样本中,由CYP2C19*2和*3等位基因突变而产生的六种基因型(*1/*1,*2/*2,*3/*3,*1/*2,*1/*3,*2/*3)均通过DNA微阵列芯片法检测出。西北地区人群中,CYP2C19*2突变常见,为52.64%(n=279),其中,纯合突变率8.87%(n=47);CYP2C19*3基因突变率为10.94%(n=58),而该位点的纯合突变罕见,仅0.19%(n=1);CYP2C19*2/*3双突变率为3.21%(n=17)。进而据基因型判断强代谢型(EM)465例(n=210+215+40),占87.74%,弱代谢型(PM)65例(n=47+1+17),占12.26%。结论本地区人群CYP2C19快代谢与慢代谢基因型比率与国内其他地区报道相近,但发生于*2和*3位点的杂合型突变率显著高于之前文献报道。其中*2位点的突变频率较*3更为常见(52.64%vs.10.94%)。提示CYP2C19基因多态性存在一定的地区差异,掌握本地区人群的基因分布频率对临床用药具有重要的指导意义。     

双管单色荧光PCR法与基因芯片法检测CYP2C19基因多态性的比较研究

目的 建立快速检测CYP2C19*2和CYP2C19*3 基因多态性的实验方法。方法 从样本库中选取100例血液样本。设计针对CYP2C19*2和CYP2C19*3基因多态性位点的特异性引物和taqman荧光探针,通过双管单色荧光PCR法检测基因型,与基因芯片法进行比较验证。结果 双管单色荧光PCR法能很好地鉴别CYP2C19*2和CYP2C19*3基因型,与基因芯片结果一致率为100%。结论 双管单色荧光PCR法能快速有效地进行CYP2C19*2和CYP2C19*3基因多态性分型,该方法期待用于临床实验室以指导个体化用药。     

1043例汉族双相情感障碍患者CYP2C19基因多态性及代谢表型分析

目的探讨汉族双相情感障碍患者CYP2C19基因多态性及代谢表型的分布,为临床合理用药提供理论参考和指导。方法以DNA微阵列技术检测1 043例双相情感障碍患者的CYP2C19基因多态性及代谢表型(快代谢型,中等代谢型,慢代谢型),用基因计数法计算基因型频率和等位基因频率,同时分析基因多态性发生频率与性别的相关性。结果所有检测对象中,CYP2C19*1/*1、*1/*2、*1/*3、*2/*2、*2/*3、*3/*3基因型的频率分别为40.6%、40.5%、5.2%、9.9%、3.4%、0.5%;CYP2C19基因快代谢型、中等代谢型、慢代谢型3种代谢表型所占比例分别为40.5%、45.8%、13.7%;不同性别之间CYP2C19基因型及代谢表型差异无统计学意义(P0.05)。结论汉族双相情感障碍人群CYP2C19基因多态性以*1/*1及*1/*2为主,681位点的变异率高于636位点,检测CYP2C19基因多态性及代谢表型有助于临床合理用药。     

基质辅助激光解吸电离飞行时间质谱技术检测药物代谢酶基因多态性平台的建立

目的 建立基于基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术检测患者体内药物代谢酶基因多态性的平台。方法 选取2013年10月～2014年6月在北京协和医院门诊就诊患者53例EDTA抗凝外周血,提取全血基因组DNA,用MALDI-TOF-MS技术检测53例患者药物代谢酶基因CYP2C9*2(rs1799853),CYP2C9*3(rs1057910),CYP2C19*2(rs4244285),CYP2C19*3(rs4986893),CYP2C19*4(rs28399504),CYP2C19*5(rs56337013)和CYP2C19*17(rs12248560)的单核苷酸多态性(SNP)位点,并用Sanger测序法进行验证。结果 用 MALDI-TOF-MS 技术可以同时完成53份样本,2个药物代谢酶基因,7个SNP位点的检测。53例患者中,发现CYP2C19*2(rs4244285)AG型25例,AA型6例,GG型22例,A等位基因频率为34.9%。CYP2C19*3(rs4986893)AG型4例,GG型49例,A等位基因频率为3.8%。CYP2C9*3(rs1057910)CA型5例,AA型48例,C等位基因频率为4.7%。未发现 CYP2C9*2(rs1799853),CYP2C19*4(rs28399504),CYP2C19*5(rs56337013)和CYP2C19*17(rs12248560)位点突变。经与Sanger测序法比较,两种检测方法结果的符合率为100%。结论 成功建立MALDI-TOF-MS技术检测药物代谢酶基因多态性的平台,该平台具有高通量、准确的特点,对个性化用药治疗具有重要的临床应用价值。     

CYP2C19基因多态性与丙戊酸血药浓度相关性及个体化用药研究

目的观察分析CYP2C19基因多态性与癫痫患者丙戊酸血药浓度的个体差异,并为临床个体化用药提供参考依据。方法选取2015年1月至2016年12月于该院就诊的癫痫患者148例,采用实时荧光聚合酶链反应(PCR)技术检测148例患者的CYP2C19基因分型,同时给予丙戊酸治疗,分析CYP2C19基因多态性与丙戊酸血药浓度的相关性。结果不同CYP2C19基因型的癫痫患者丙戊酸血药浓度存在很大的个体差异,其中148例癫痫患者等位基因和基因型分布频率比较,差异无统计学意义(P0.05);CYP2C19*2位点的AA基因型服药2、4、8h后丙戊酸血药浓度水平显著高于CYP2C19*2位点GG基因型,差异有统计学意义(P0.05);CYP2C19*3位点AA基因型服药8h后丙戊酸血药浓度水平显著高于8h后丙戊酸血药浓度水平CYP2C19*3位点GG基因型,差异有统计学意义(P0.05);其他基因型丙戊酸血药浓度水平比较,差异无统计学意义(P0.05)。结论癫痫患者的CYP2C19基因具有多态性,且与服用丙戊酸治疗的癫痫患者血液浓度具有相关性。因此,在采用丙戊酸治疗癫痫患者时,可检测患者CYP2C19基因分型,以指导癫痫患者的临床个体化治疗。     

四川地区冠心病患者CYP2C19基因多态性检测

目的检测四川地区冠心病患者CYP2C19基因的多态性,分析基因型与性别、年龄及临床分型之间的相关性。方法采用DNA微阵列芯片法检测131例被确诊为冠心病患者的CYP2C19基因型,结合患者的性别、年龄、临床分型进行分组,探讨这些因素与CYP2C19基因多态性的相关性。结果 131例冠心病患者中,有5种基因型被检测到,分别为CYP2C19*1/*1(636GG,681GG)、CYP2C19*1/*2(636GG,681GA)、CYP2C19*1/*3(636GA,681GG)、CYP2C19*2/*2(636GG,681AA)、CYP2C19*2/*3(636GA,681GA)。分布频率分别为53例(40.5%)、54例(41.2%)、6例(4.6%)、13例(9.9%)、5例(3.8%),而CYP2C19*3/*3(636AA,681GG)未被检测到。不同性别冠心病患者CYP2C19的基因型、等位基因频率差异无统计学意义(P0.05);不同性别的代谢型患者,其CYP2C19的基因型分布差异具有统计学意义(χ~2=6.527,P=0.038)。不同年龄和临床分型冠心病患者CYP2C19的基因型、等位基因频率、代谢型分布差异无统计学意义(P0.05)。结论冠心病患者的CYP2C19基因型分布、等位基因频率分布与性别、年龄和疾病临床分型无关。     

潮汕地区人群CYP2C9基因多态性研究

目的了解细胞色素酶P450家族CYP2C9基因多态性在潮汕地区人群分布的情况,以及基因型分布和基因突变率。方法采用DNA测序法检测204例当地健康志愿者CYP2C9*2和CYP2C9*3的基因多态性。结果204例志愿者CYP2C9基因型检测有184例为*1/*1型,19例为*1/*3型,1例为*3/*3型,频率分别为90.19%、9.31%和0.50%,未发现*1/*2、*2/*2和*2/*3型突变;CYP2C9*3的突变率在男女之间差异有统计学意义(P0.05)。结论 CYP2C9基因在潮汕地区的多态性分布和性别有关,根据基因型的差异进行个体化用药有利于临床治疗。     

华南地区汉族人群心脑血管相关基因多态性分析

目的了解心脑血管相关基因细胞色素P450酶(CYP)2C19、CYP2C9、维生素K环氧化物还原酶复合体1(VKORC1)和乙醛脱氢酶2(ALDH2)基因的单核苷酸多态性频率在华南地区汉族人群分布的情况。方法采用芯片技术对252例心脑血管疾病患者的CYP2C19、ALDH2、CYP2C9以及VKORC1位点基因多态性进行检测,并统计男女间基因型频率的差异。结果 CYP2C19的野生型*1/*1占42.5%,突变型*1/*2占36.9%,*1/*3占7.9%,*2/*2占9.9%,*2/*3占1.6%,*3/*3占1.2%,在男女人群间各类基因型的发生率差异无统计学意义(P0.05);CYP2C9只检测到2种等位基因CYP2C9*1/*1和CYP2C9*1/*3,男性基因频率分别为98.6%和1.4%,女性基因频率分别为90.1%和9.9%,在男女人群间的分布差异有统计学意义(P0.05);VKORC1有3种基因型AA、AG和GG,频率分别为80.6%、18.7%和0.8%,在男女人群间的分布差异无统计学意义(P0.05);ALDH2有3种基因型GG、GL和LL型,频率分别为55.2%、37.3%和7.5%,在男女人群间的分布差异无统计学意义(P0.05)。结论 CYP2C9的多态性分布与性别有关,CYP2C19、VKORC1和ALDH2的多态性分布与性别无关。     

DNA测序法检测CYP2C19基因多态性指导氯吡格雷用药的临床价值

目的评估DNA测序法检测细胞色素氧化酶P450 2C19(CYP2C19)多态性以指导氯吡格雷临床用药。方法选取2010年11月至2013年11月武汉大学人民医院心血管内科诊断为急性冠状动脉综合征,并首次接受经皮冠状动脉介入治疗术(PCI)的140例患者为研究对象,随机分为试验组及对照组各70例。试验组采集外周血标本并提取基因组DNA,根据已知的CYP2C19基因序列,设计针对CYP2C19*2、CYP2C19*3位点的特异性引物,聚合酶链反应扩增包含CYP2C19基因多态性位点的DNA片段;并以DNA测序法进行鉴定,分析不同CYP2C19基因型在试验组患者中的分布频率。对照组患者使用氯吡格雷药物但未检测相关基因。比较两组患者随访过程中发生冠状动脉血栓事件的差异。结果 DNA测序结果显示,试验组中共有4种基因型组合被检测到:*1/*1(636GG,681GG)、*1/*2(636GG,681GA)、*2/*2(636GG,681AA)、*1/*3(636GA,681GG),分布频率分别为33例(47.1%)、29例(41.4%)、4例(5.7%)、4例(5.7%),而*3/*3(636AA,681GG)和*2/*3(636GA,681GA)未检测到。经基因型指导氯吡格雷用药的试验组患者发生支架冠状动脉血栓事件的比例为0.0%,明显低于对照组的7.1%,差异有统计学意义(P0.05)。结论基因型指导氯吡格雷用药有助于患者用药剂量的调整,可降低冠状动脉血栓事件的发生率。     

等位基因特异性PCR检测CYP3A5和MDR-1基因多态性方法的建立及临床应用

目的用等位基因特异性PCR(ARMS-PCR)检测细胞色素P450酶CYP3A5(A6986G)和多药耐药基因MDR-1(C3435T)基因多态性,探讨其与肾移植受者他克莫司(Tac)血药浓度和剂量比的相关性。方法根据CYP3A5(A6986G)和MDR-1(C3435T)基因多态性位点分别设计ARMS-PCR引物,分析72例肾移植受者外周血基因组DNA中该2个基因位点的多态性,同时以DNA测序法为金标准进行验证。化学发光微粒子免疫分析法测定肾移植受者血Tac浓度,并比较术后1个月时不同基因型受者之间血Tac浓度、Tac剂量/Tac用量的差异。结果建立的ARMS-PCR法与DNA测序法检测符合率为100%。72例肾移植受者中,CYP3A5*1/*1、*1/*3和*3/*3基因型的发生频率分别为18.1%、31.9%和50.0%,MDR-1 C/C、C/T和T/T基因型的发生频率分别为27.8%、58.3%和13.9%。此外,不同CYP3A5基因型肾移植受者的血Tac浓度(P=0.014)和Tac浓度/Tac用量(P=0.019)均存在明显差异,进一步两两比较发现,CYP3A5*3/*3基因型受者血Tac浓度/Tac用量明显高于*1/*1和*1/*3基因型(P均0.05)。结论成功建立检测CYP3A5和MDR-1基因多态性的ARMS-PCR法,肾移植受者CYP3A5*3基因多态性与Tac的药代动力学明显相关。     

CYP2C9 and CYP2C19 genetic polymorphisms: frequencies in the south Indian population

The aim of the study was to establish the frequencies of CYP2C9*1, *2, *3 and CYP2C19*1, *2 and *3 in the south Indian population and to compare them with the inter-racial distribution of the CYP2C9 and CYP2C19 genetic polymorphisms. Genotyping analyses of CYP2C9 and CYP2C19 were conducted in unrelated, healthy volunteers from the three south Indian states of Andhra Pradesh, Karnataka and Kerala, by the polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP). The allele frequencies of the populations of these three states were then pooled with our previous genotyping data of Tamilians (also in south India), to arrive at the distribution of CYP2C9 and CYP2C19 alleles in the south Indian population. Frequencies of CYP2C9 and CYP2C19 alleles and genotypes among various populations were compared using the two-tailed Fisher's exact test. The frequencies of CYP2C9*1, *2 and *3 in the south Indian population were 0.88 (95% CI 0.85-0.91), 0.04 (95% CI 0.02-0.06) and 0.08 (95% CI 0.06-0.11), respectively. The frequencies of CYP2C9 genotypes *1/*1, *1/*2, *1/*3, *2/*2, *2/*3 and *3/*3 were 0.78 (95% CI 0.74-0.82), 0.05 (95% CI 0.03-0.07), 0.15 (95% CI 0.12-0.18), 0.01 (95% CI 0.0-0.02), 0.01 (95% CI 0.0-0.02) and 0.0, respectively. CYP2C19*1, *2 and *3 frequencies were 0.64 (95% CI 0.60-0.68), 0.35 (95% CI 0.31-0.39) and 0.01 (95% CI 0.0-0.03), respectively. As a result of a significant heterogeneity, the data on CYP2C19 genotype frequencies were not pooled. The frequency of CYP2C9*2 mutant alleles in south Indians was higher than in Chinese and Caucasians, while CYP2C9*3 was similar to Caucasians. CYP2C19*2 was higher than in other major populations reported so far. The relatively high CYP2C19 poor-metabolizer genotype frequency of 12.6% indicates that over 28 million south Indians are poor metabolizers of CYP2C19 substrates.     

Interindividual variability in ibuprofen pharmacokinetics is related to interaction of cytochrome P450 2C8 and 2C9 amino acid polymorphisms

OBJECTIVE: Our objective was to identify genetic factors related to interindividual variability in the pharmacokinetics of ibuprofen and its enantiomers. METHODS: The time course for ibuprofen plasma concentration was measured by HPLC in 130 healthy individuals who received a single oral dose of 400 mg racemic ibuprofen. Genomic deoxyribonucleic acid was analyzed for common mutations at CYP2C8 and CYP2C9 genes that cause amino acid substitutions. RESULTS: Ibuprofen clearance values were 4.04 L/h (95% confidence interval [CI], 3.61-4.47 L/h), 2.79 L/h (95% CI, 2.07-3.52 L/h), and 0.40 L/h (95% CI, 0.37-0.43 L/h) for carriers of CYP2C8 genotypes *1/*1, *1/*3, and *3/*3, respectively, and 4.43 L/h (95% CI, 3.94-4.92 L/h), 3.26 L/h (95% CI, 2.53-3.99 L/h), 2.91 L/h (95% CI, 1.52-4.30 L/h), 2.05 L/h (95% CI, 0-6.37 L/h), 1.83 L/h (95% CI, 1.24-2.41 L/h), and 1.13 L/h (95% CI, 0.58-1.66 L/h) for carriers of the CYP2C9 genotypes *1/*1, *1/*2, *1/*3, *2/*2, *2/*3, and *3/*3, respectively. The P values for comparison across nonmutated, heterozygous, and homozygous genotypes were as follows: P <.001 for CYP2C8*3, P <.005 for CYP2C9*2, and P <.001 for CYP2C9*3. The main genetic factor for reduced clearance of R-(-)-ibuprofen is the CYP2C8*3 allele, whereas the clearance for S-(+)-ibuprofen is influenced by CYP2C8*3 and CYP2C9*3 alleles to a similar extent. The CYP2C9*2 allele was associated with low clearance only when it was present in combination with the CYP2C8*3 allele. As compared with individuals with no mutations, individuals with the common genotype CYP2C8*1/*3 plus CYP2C9*1/*2 (19% of the population) displayed decreased ibuprofen clearance (mean, 65% [95% CI, 42%-89%]; P <.001). Individuals homozygous or double-heterozygous for CYP2C8*3 and CYP2C9*3 variant alleles (8% of the population) had extremely low ibuprofen clearance rates, with values ranging from 7% to 27% of the mean clearance rates among noncarriers of mutations (P <.001). No enantiospecific reduction of ibuprofen clearance was observed. CONCLUSION: Low ibuprofen clearance occurs in a substantial proportion of healthy subjects, is not enantiospecific, and is strongly linked to CYP2C8 and CYP2C9 polymorphisms.     

Enantiospecific effects of cytochrome P450 2C9 amino acid variants on ibuprofen pharmacokinetics and on the inhibition of cyclooxygenases 1 and 2

OBJECTIVE: According to in vitro data, the polymorphic cytochrome P450 enzyme 2C9 (CYP2C9) may be the major S-ibuprofen hydroxylase. In humans, there are 2 variants of CYP2C9 with a high population frequency. We studied their impact on ibuprofen pharmacokinetics and on the inhibition of cyclooxygenases 1 and 2. METHODS: Kinetics of an oral dose of 600 mg racemic ibuprofen were studied in 21 healthy volunteers with all combinations of the CYP2C9 variants *2 (arginine144cysteine) and *3 (isoleucine359leucine). Blood concentrations of racemic ibuprofen and of S-(+)-ibuprofen and R-(-)-ibuprofen were measured by HPLC, and thromboxane B(2) and prostaglandin E(2) were measured with use of an enzyme immunoassay. Data were evaluated with a population pharmacokinetic model that integrated pharmacogenetic information. RESULTS: The pharmacokinetics of racemic and of S-ibuprofen depended on the CYP2C9 isoleucine359leucine amino acid polymorphism: population mean S-ibuprofen clearances were 3.25 L/h (95% confidence interval [CI], 2.84 to 3.73), 2.38 L/h (95% CI, 2.09 to 2.73), and 1.52 L/h (95% CI, 1.33 to 1.74) in carriers of the CYP2C9 genotypes *1/*1, *1/*3, and *3/*3, respectively. The CYP2C9 variant *2 exhibited no significant effect. Ex vivo formation of thromboxane B(2), reflecting cyclooxygenase type 1 inhibition, depended significantly on the CYP2C9 polymorphism. The maximal inhibition of thromboxane B(2) formation and the area under the effect-time curve were larger in carriers of the slow CYP2C9 genotypes *1/*3, *2/*3, and *3/*3 than in *1/*1 carriers; the same trend was observed for prostaglandin E(2), reflecting cyclooxygenase type 2 inhibition. CONCLUSIONS: The reduced S-ibuprofen total clearance accompanied by increased pharmacodynamic activity may have medical impact in patients receiving ibuprofen.     

武汉地区冠心病患者氯吡格雷药物代谢相关基因CYP2C19的多态性分布分析

目的 探讨武汉地区冠心病介入患者氯吡格雷代谢相关基因CYP2C19多态性的分布。方法 选取2014年1月～12月武汉大学人民医院心内科进行介入治疗(PCI)的316例冠心病患者作为研究对象。通过基因芯片法检测氯吡格雷代谢相关的CYP2C19*1,*2,*3基因,并将患者按CYP2C19基因型别分为不同代谢类型:强代谢型(*1/*1),中间代谢型(*1/*2,*1/*3),弱代谢型(*2/*2,*3/*3,*2/*3)。结果 根据CYP2C19基因多态性位点功能代谢分型,携带CYP2C19*1的强代谢型(*1/*1)占43.4%,携带CYP2C19*2或*3的中间代谢型(*1/*2和*1/*3)及弱代谢型(*2/*2,*2/*3和*3/*3)分别占42.4%,14.2%。不同性别在CYP2C19基因分型上差异无统计学意义。结论 武汉地区冠心病介入患者中分布有较多的CYP2C19氯吡格雷代谢功能缺失基因。     

Pharmacokinetics of the new proton pump inhibitor ilaprazole in Chinese healthy subjects in relation to CYP3A5 and CYP2C19 genotypes

BACKGROUND: PPIs are widely used in peptic diseases, and this paper is to investigate the kinetic characteristics of a new PPI ilaprazole in Chinese healthy subjects and the association with CYP3A5 and CYP2C19 polymorphisms. METHODS: 21 subjects were selected and treated with 10mg ilaprazole according to their CYP3A5*3 genotypes (including 7 of CYP3A5*1/*1, 7 of *1/*3, and 7 of *3/*3). The plasma concentrations of ilaprazole and its metabolites were monitored by LC-MS/MS method. RESULTS: The C(max), AUC((0-6)), AUC((0-48)) and AUC((0-infinity)) of ilaprazole were all significantly different across the 3 CYP3A5 genotypes (including 4 of CYP3A5*1/*1, 4 of *1/*3, 3 of *3/*3; P<0.05) in CYP2C19 wild-type subjects (CYP2C19 wt/wts), similar variety of C(max) and AUC((0-6)) among CYP3A5 genotypes (including 3 of CYP3A5*1/*1, 3 of *1/*3, 4 of *3/*3; P<0.05) were also observed in CYP2C19 heterozygous subjects (CYP2C19 wt/mts). The sulfoxidation metabolic index (measure of collective CYP3A activity) indicates that the CYP3A5*1/*1, (high-expressers), *1/*3, (low-expressers), and *3/*3 (no-expressers) groups have medium, lowest and highest activities on ilaprazole metabolism, inconsistent with genotype-based CYP3A5 enzymatic activity. Further analysis showed no correlation between ilaprazole metabolism and CYP2C19 genotypes, evidenced by that the AUC((0-infinity)) of ilaprazole from either CYP3A5*1/*1 or CYP3A5*1/*3 groups was much higher in CYP2C19 wt/wts (n=4) than that in CYP2C19 wt/mts (n=3) (P<0.001), but the C(max) and AUC((0-6)) of ilaprazole from CYP3A5*3/*3 groups, were significantly lower in CYP2C19 wt/wts (n=3) compared to CYP2C19 wt/mts (n=4) (P<0.01). CONCLUSIONS: There was no demonstrated relationship between ilaprazole metabolism and CYP3A5 polymorphisms.     

DNA微阵列芯片法与直接测序法检测CYP2C19基因型的比较研究

目的 用DNA微阵列芯片法和直接测序法两种方法检测氯吡格雷相关基因CYP2C19的突变情况,并进行比较分析。同时将基因型检测结果与临床资料进行分析,初步探讨CYP2C19基因型检测在氯吡格雷用药治疗中的临床意义。方法收集180例诊断为急性冠脉综合症并首次接受经皮冠状动脉介入治疗术(PCI)的全血标本。其中90例患者术后服用氯吡格雷之前,采用DNA微阵列芯片法和DNA直接测序法检测CYP2C19突变位点,确定其基因型。另外90例对照组患者使用氯吡格雷药物但不检测CYP2C19基因型。分析两组患者随访过程中发生冠脉血栓事件的差异。结果 ①两种检测方法准确度比较:在试验组90例患者中,DNA微阵列芯片法和DNA测序法均检出4种基因型组合:*1/*1(636GG,681GG),*1/*2(636GG,681GA),*2/*2(636GG,681AA)和*1/*3(636GA,681GG),分布频率分别为44例(48.9%),36例(40%),5例(5.6%)和5例(5.6%),而*3/*3(636AA,681GG)和*2/*3(636GA,681GA)未检测到。两种方法的准确度完全一致。②将检测结果与临床资料进行相关性分析:经CYP2C19基因型指导氯吡格雷用药的患者发生支架冠脉血栓事件的比例(0%)明显低于对照组(3.33%,P<0.05)。结论①DNA微阵列芯片法灵敏度和准确度与DNA直接测序法(金标准方法)检测相符率为100%。②DNA微阵列芯片法是一种快速、准确发现CYP2C19突变位点,鉴别CYP2C19基因型的检测方法。③检测结果与临床相关性分析结果显示基因型指导氯吡格雷用药有助于患者用药剂量的调整,或选用其它抗凝血药物,从而可降低冠脉血栓事件的发生率。     

Pharmacokinetics and pharmacodynamics of rosiglitazone in relation to CYP2C8 genotype

OBJECTIVES: Rosiglitazone is metabolically inactivated predominantly via the cytochrome P450 (CYP) enzyme CYP2C8. The functional impact of the CYP2C8*3 allele coding for the Arg139Lys and Lys399Arg amino acid substitutions is controversial. The purpose of this was to clarify the role of this polymorphism with regard to the pharmacokinetics and clinical effects of rosiglitazone. METHODS: From a large sample of healthy volunteers, 14 carriers of the CYP2C8*1/*1 allele, 13 carriers of the *1/*3 allele, and 4 carriers the *3/*3 allele were selected for a clinical study. Rosiglitazone (8 mg) single-dose and multiple-dose pharmacokinetics and its effects on glucose level and body weight were monitored. Plasma and urine concentrations of rosiglitazone and desmethylrosiglitazone were measured, and kinetics was analyzed by noncompartmental and population-kinetic compartmental methods. RESULTS: Mean total clearance values were 0.033 L x h(-1) x kg(-1) (95% confidence interval [CI], 0.030-0.037 L x h(-1) x kg(-1)), 0.038 L x h(-1) x kg(-1) (95% CI, 0.033-0.044 L x h(-1) x kg(-1)), and 0.046 L x h(-1) x kg(-1) (95% CI, 0.033-0.058 L x h(-1) x kg(-1)) in carriers of CYP2C8 genotypes *1/*1, *1/*3, and *3/*3, respectively, on day 1 (P = .02, ANOVA [F test]). Rosiglitazone kinetics could be adequately described by a 1-compartmental model with first-order absorption. Besides CYP2C8 genotype, body weight was a significant covariate (P < .001, log-likelihood ratio test). Elimination half-lives were 4.3, 3.5, and 2.9 hours in CYP2C8*1/*1, *1/*3, and *3/*3 carriers, respectively. Clearance of desmethylrosiglitazone was also higher in CYP2C8*3 allele carriers, with mean values of 1.96 L/h (95% CI, 1.42-2.69 L/h), 2.22 L/h (95% CI, 1.61-3.04 L/h), and 2.47 L/h (95% CI, 1.80-3.39 L/h), respectively (P = .03). The plasma glucose area under the concentration curve was significantly lower after 14 days of taking rosiglitazone compared with day 1 (P = .01, paired t test), but no relationship of the glucose-lowering effect of rosiglitazone with CYP2C8 genotype was observed. CONCLUSIONS: This study showed that the CYP2C8*3 allele confers higher in vivo metabolic capacity than the wild-type CYP2C8*1 allele but the pharmacokinetic differences resulting from CYP2C8*3 were quantitatively moderate.     

CYP2C9, but not CYP2C19, polymorphisms affect the pharmacokinetics and pharmacodynamics of glyburide in Chinese subjects

BACKGROUND: Although cytochrome P450 (CYP) 2C9 was thought to be the main pathway for glyburide (INN, glibenclamide) metabolism in vivo, studies in vitro indicated that CYP2C19 had a more dominant effect. This study investigated the relative influence of CYP2C9 and CYP2C19 genotypes on the pharmacokinetics and pharmacodynamics of glyburide in Chinese subjects. METHODS: Three groups of healthy male Chinese subjects (n=6 per group) were enrolled, as follows: group I, CYP2C9*1/*1 and CYP2C19 extensive metabolizers (EMs); group II, CYP2C9*1/*1 and CYP2C19 poor metabolizers (PMs); and group III, CYP2C9*1/*3 and CYP2C19 EMs. Subjects received single oral doses of 5 mg glyburide. Multiple blood samples were collected, and the plasma glyburide concentrations were determined by an HPLC method. The plasma glucose and insulin concentrations were also measured up to 2 hours after dosing. RESULTS: No significant differences in glyburide pharmacokinetics were observed between CYP2C19 EM and PM subjects who had the CYP2C9*1/*1 genotype (group I versus group II). Their respective values for area under the plasma concentration-time curve from time 0 to infinity (AUC0-infinity) and elimination half-life (t1/2) were 0.46+/-0.13 microg.h/mL versus 0.57+/- 0.11 microg.h/mL (P=.569) and 2.09+/-0.22 hours versus 2.24+/- 0.27 hours (P=.721). However, significant increases in AUC(0-infinity) (125% and 82%; P=.008 and .024, respectively) and t1/2 (71% and 60%; P=.003 and .007, respectively) were observed when CYP2C9*1/*3 subjects (group III) were compared with CYP2C9*1/*1 subjects in group I or II. Blood glucose reductions at 2 hours after dosing were 41.8%, 23.9%, and 27.7% in groups I, II, and III, respectively (P=.029), and hypoglycemia developed in 3 of 6 CYP2C9*1/*3 carriers and 2 of 12 CYP2C9*1/*1 carriers. CONCLUSION: CYP2C9, but not CYP2C19, polymorphism appears to exert a dominant influence on glyburide pharmacokinetics and pharmacodynamics in vivo. Further studies in diabetic patients with long-term dosing are warranted to confirm these findings.     

东莞地区汉族冠心病患者CYP2C19基因多态性分布特征分析

目的探讨东莞地区汉族冠心病患者CYP2C19基因多态性分布特征。方法选取该院2016年9月至2019年1月收治的800例汉族冠心病患者作为研究对象,采用等位基因特异PCR技术对患者CYP2C19基因多态性进行检测,了解CYP2C19基因多态性分布情况,并根据患者的年龄、性别进行分组分析。结果 CYP2C19基因型中*1/*1出现频率为38.50%,*1/*2频率为40.00%,*1/*3频率为6.38%,*2/*2频率为11.75%,*2/*3频率为3.00%,*3/*3频率为0.37%;其中快代谢型为38.50%、中代谢型为46.38%、慢代谢型为15.12%;等位基因分布中CYP2C19*1占61.69%,CYP2C19*2占33.25%,CYP2C19*3占5.06%。不同年龄、性别、患者CYP2C19各基因多态性分布虽存在一定的差异,但差异无统计学意义(P>0.05)。结论东莞地区汉族人群冠心病患者CYP2C19基因型中以快代谢型与中代谢型为主,基因分布以CYP2C19*1占比较高,不同性别、年龄患者CYP2C19基因型间差异不明显。